In vivo immunofluorescence to diagnose herpes simplex virus keratitis in mice

BACKGROUND/AIMS: Herpes simplex virus keratitis (HSK) is the most common cause of corneal blindness in the Western world. Delay in the treatment of HSK can lead to a more significant corneal scar and topical steroid treatment in unsuspected active HSK can lead to corneal melting. Current culture techniques for herpes simplex virus (HSV) take several days and commercially available HSV laboratory based diagnostic techniques such as Herpchek vary in sensitivity. This study was conducted to assess the viability of a new, quicker, and simpler method to diagnose HSK. METHODS: Direct immunofluorescence was used in vivo in a masked study to diagnose HSK in mice using a standard slit lamp with cobalt blue illumination. Murine monoclonal fluorescently labelled antibody was applied to the cornea for 10 or 20 minutes and then washed off with phosphate buffered solution. Mice with HSK were stained with either fluorescently labelled monoclonal antibody against HSV or fluorescently labelled monoclonal antibody against cytomegalovirus. Mice with corneal abrasions of non-viral origin were given fluorescently labelled monoclonal antibody against HSV. RESULTS: Fluorescence was seen only in the mice with HSK given fluorescently labelled monoclonal antibody against HSV. This observation was confirmed upon microscopic immunofluorescent imaging of the corneal epithelial sheets. CONCLUSION: In vivo immunofluorescence may be useful in the clinical diagnosis of HSK.     

Antiviral therapy for recurrent herpes simplex reconsidered

The possible association of leukotriene B4 (LTB4)-like activity with the development of active rheumatoid arthritis was studied in 25 children with the disease and in 15 normal subjects. Serum LTB4-like activity was found to be significantly higher in the active stage of the disease when compared with the values obtained from patients during the inactive stage and from healthy children. No correlation was found between LTB4-activity and other laboratory parameters, e.g. haemoglobin level, white cell count and erythrocyte sedimentation rate.     

A herpes simplex virus DNA polymerase mutation that specifically attenuates neurovirulence in mice

Herpes simplex virus can infect the mammalian brain causing lethal encephalitis (neurovirulence). Previously, herpes simplex virus mutants that are attenuated for neurovirulence have exhibited defects in replication in brain and/or blocks to replication in neuronal cells. We investigated the attenuation of neurovirulence of mutant PAAr5, which exhibits resistance to antiviral drugs due to altered viral DNA polymerase. Following intracerebral inoculation of 7-week-old CD1 mice, PAAr5 was 30-fold attenuated for neurovirulence compared to its wild-type parent. A drug-sensitive virus derived by marker rescue with DNA polymerase gene sequences exhibited neurovirulence that was essentially indistinguishable from that of wild-type virus, demonstrating that attenuation was due to a polymerase mutation. PAAr5 replicated in brain similarly to wild-type virus unlike another polymerase mutant, 615.8, that exhibited a similar degree of attenuation. The attenuation of PAAr5 was not associated with altered particle to PFU ratios nor with any obvious reductions in viral antigen expression in neurons, spread, histopathology, or TUNEL staining suggestive of apoptotic cells. Thus PAAr5 differs from other mutants that are attenuated for neurovirulence. Understanding how a polymerase mutation specifically attenuates neurovirulence may shed light on how herpes simplex virus can cause lethal encephalitis.     

Stromal keratitis induced by a unique clinical isolate of herpes simplex virus type 1

The antibody responses of 65 volunteers receiving an i.d. regimen (0.1 ml given at two sites on days 0, 3, 7 and 0.1 ml given at one site on days 30 and 90) were compared with the control group of 35 volunteers receiving the standard i.m. regimen. By day 14, seroconversion was observed in all vaccinees in both groups. Geometric Mean Titers remained higher than 0.5 IU/ml throughout the study period. At the end of the observation period on day 365, antibodies persisted in all subjects. The multisite i.d. PCEC regimen has been proved as immunogenic as the standard i.m. regimen. Both regimens were well tolerated. Thus, it would be the effective and cheapest available rabies post-exposure treatment using tissue culture vaccine.     

In vivo production of cytokines and beta (C-C) chemokines in human recurrent herpes simplex lesions--do herpes simplex virus-infected keratinocytes contribute to their production?

Recurrent human herpes simplex lesions are infiltrated by macrophages and CD4 and CD8 lymphocytes, which secrete cytokines and chemokines. Vesicle fluid was examined by ELISA for the presence of cytokines and beta (C-C) chemokines. On the first day of the lesion, high concentrations of interleukin (IL)-1beta, and IL-6, moderate concentrations of IL-1alpha and IL-10, and low concentrations of IL-12 and beta chemokines were found; levels of macrophage inflammatory protein (MIP)-1beta were significantly higher than levels of MIP-1alpha and RANTES. At day 3, the concentrations of IL-1beta, IL-6, and MIP-1beta were lower, whereas the levels of IL-10, IL-12, and MIP-1alpha remained similar, and the level of tumor necrosis factor-alpha was now detectable. Herpes simplex virus infection of keratinocytes in vitro stimulated production of beta chemokines followed by IL-12 and then IL-10, IL-1alpha, IL-1beta, and IL-6, indicating a potential role for these events in early recruitment, activation, and interferon-gamma production of CD4 cells in herpetic lesions.     

Cytomegalovirus and herpes simplex virus in pathogenesis and progression of native arteriosclerosis and recurrent stenosis after intervention]

Recently, the number of patients with atherosclerosis has been rapidly increased in Japan. However, there are many kinds of resolving problems for management of those patients. Here history and current topics of coronary artery bypass grafting (CABG) in Japan are clearly documented. That is, CABG could be done in 48,612 cases during the past 5 years (1991-1995). And its mortality rate was 2.3% approximately. On the other hand, 12,025 underwent CABG in 1996 in Japan. Further several kinds of operative methods including MIDCAB have been carried out with the number of the patients with ischemic heart diseases. Excellent results have been obtained respectively.     

Protective vaccination against primary and recurrent disease caused by herpes simplex virus (HSV) type 2 using a genetically disabled HSV-1

The vaccine potential of a mutant herpes simplex virus (HSV) type 1, with a deletion in the glycoprotein H (gH) gene, was evaluated. The virus requires a gH-expressing cell line for multi-cycle growth but can complete a single cycle of infection in noncomplementing cells. Such viruses, termed DISC (disabled infectious single cycle) viruses, should be safe, yet still able to stimulate humoral and cell-mediated responses against a broad range of virus antigens in vaccinated hosts. Prophylactic vaccination of guinea pigs with DISC HSV-1, by ear scarification or direct infection of the vaginal mucosa, afforded a high degree of protection against HSV-2-induced primary genital disease and reduced significantly the frequency of subsequent disease recurrence. There was also a trend toward reduced recurrence following therapeutic vaccination of animals already infected with HSV-2. DISC HSV vaccination, therefore, offers an effective route for control of HSV disease.     

Efficacy of 5,6-dihydro-5-azathymidine against cutaneous herpes simplex virus in hairless mice

5,6-Dihydro-5-azathymidine, administered subcutaneously, was active both prophylactically and therapeutically against cutaneous herpesvirus infection of hairless mice. Activity was comparable to that obtained with adenine arabinoside.     

Interleukin-18 protects mice against acute herpes simplex virus type 1 infection

We examined the effects of interleukin-18 (IL-18) in a mouse model of acute intraperitoneal infection with herpes simplex virus type 1 (HSV-1). Four days of treatment with IL-18 (from 2 days before infection to 1 day after infection) improved the survival rate of BALB/c, BALB/c nude, and BALB/c SCID mice, suggesting innate immunity. One day after infection, HSV-1 titers were higher in the peritoneal washing fluid of control BALB/c mice than in that of IL-18-treated mice. A genetic deficiency of gamma interferon (IFN-gamma), however, diminished the survival rate and the inhibition of HSV-1 growth at the injection site in the mice. Anti-asialo GM1 treatment had no influence on the protective effect of IL-18 in infected mice. IL-18 augmented IFN-gamma release in vitro by peritoneal cells from uninfected mice, while no appreciable IFN-gamma production was found in uninfected mice administered IL-18. Although IFN-gamma has the ability to induce nitric oxide (NO) production by various types of cells, administration of the NO synthase inhibitor NG-monomethyl-L-arginine resulted in superficial loss of the improved survival, but there was no influence on the inhibition of HSV-1 replication at the injection site in IL-18-treated mice. Based on these results, we propose that IFN-gamma produced before HSV-1 infection plays a key role as one of the IL-18-promoted protection mechanisms and that neither NK cells nor NO plays this role.     

Diagnosis of herpes simplex diseases

The diagnosis of herpes simplex diseases will be made on ground of the typical clinical symptoms in more than 90% of all cases. A very easy additional procedure is the Tzanck test. If possible, immunofluorescent microscopy and electron microscopy of negatively stained virus particles may be helpful. Remaining questions should be solved by further virological examination in special laboratories. Information about sampling and shipment of the diagnostic material (vesicle fluid, swabs from mouth and genital ulcers, blood etc.) should be obtained prior to collection.     

Immunotherapy of recurrent genital herpes with recombinant herpes simplex virus type 2 glycoproteins D and B: results of a placebo-controlled vaccine trial

To determine the safety, immunogenicity, and efficacy of a recombinant herpes simplex virus type 2 glycoprotein D and B vaccine in the treatment of recurrent genital herpes, a randomized, placebo-controlled trial was held at two referral centers. Healthy patients with 4-14 recurrences per year received injections of both glycoproteins in MF59 adjuvant or of MF59 alone at 0, 2, 12, and 14 months. For 18 study months, the rate and number of recurrences, the duration and severity of the first confirmed recurrence, vaccine immunogenicity, and rates of local and systemic reactions were determined. The monthly rate of recurrences was not significantly improved, but the duration and severity of the first study outbreak was reduced significantly by vaccination. Glycoprotein-specific and neutralizing antibodies were boosted by vaccination for the duration of the study. This vaccine is safe and immunogenic and ameliorated an observed first postvaccination genital recurrence, but it does not reduce recurrence frequency.     

Para-aminobenzoic acid in therapy of experimental keratitis caused by herpes simplex virus in rabbits: the therapeutic effect and decrease of infectious titer

Immediate and long-term outcomes of repeated and multistage operations were analyzed in 190 patients with pulmonary tuberculosis. The aspects of work rehabilitation were studied in these patients. The efficiency of repeated and stage surgical interventions in patients both with disseminated and complicated types of tuberculosis and with pleural empyemas and in those with uncomplicated postoperative disease is 87.5 and 92.2%, respectively. The long-term results indicated that the complete clinical effect preserved in 79.2% of patients. Work rehabilitation was achieved in 64.8% of the examinees in the long-term postoperative period. The use of repeated and stage surgical interventions in patients with pulmonary tuberculosis may rehabilitate a rather large proportion of those operated on, assuming a great socioeconomic significance.     

Cytotoxic T lymphocyte activity after intravitreal inoculation of herpes simplex virus type I in mice

After inoculating mice with herpes simplex virus (HSV) by a corneal or intravitreal route, cytotoxic T lymphocyte (CTL) activity in the cervical lymph nodes and spleen was assayed. The spleen and cervical lymph nodes were removed at various points till 2 weeks after inoculation, and CTL activity was assayed in a groups: (A) mice intravitreally inoculated with HSV, and (B) mice with topical application of HSV. The reactivity of delayed type sensitivity was determined by the thickness of the mouse ear pinna on the 6th day in both groups. CTL activity in the spleen was at the same level in both groups. Up to 10 days after inoculation CTL activity in the cervical lymph nodes in group (A) was lower than in group (B). The reactivity of delayed type sensitivity in group (A) was lower than in group (B). These results indicate that an anterior chamber associated immune deviation (ACAID)-like phenomenon occurred after HSV inoculation into the vitreous cavity.     

Treatment of acute herpes simplex encephalitis

Combined subjective and objective nutritional assessment was performed on admission in 127 patients with oral and maxillofacial malignancies. On the basis of the nutritional assessment result, three typical nutritional parameters-body weight (BW) (X1), mid-upper-arm circumference (MAC) (X2), and hand grip strength (HGS) (X3) were used to establish a new nutritional assessment method that was developed by a computer-based discriminant analysis. The established model was as follows: Y1 = -126 + 1.09X1 + 1.34X2 + 0.23X3; Y2 = -95.63 + 0.96X1 + 1.17X2 + 0.19X3. In the model Y1 was regarded as good nutrition and Y2 as malnutrition. The larger value of Y stood for the patient's nutritional status (i.e. Y1 > Y2, well nourished; Y1 < Y2 malnourished). The new nutritional assessment correlates well with the combined subjective and objective nutritional assessment with a total agreement rate of 88.2%. With its simplicity and accuracy, the new nutritional assessment deserves a wide application in clinical situation.